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罗翠平

作品数:5 被引量:8H指数:1
供职机构:武汉工业学院生物与制药工程学院更多>>
发文基金:湖北省教育厅优秀中青年人才项目博士科研启动基金更多>>
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cry基因保守序列克隆及其在大肠杆菌Rosetta(DE3)中的表达被引量:1
2012年
[目的]对cry基因保守序列进行克隆,并使其在大肠杆菌Rosetta(DE3)中进行表达。[方法]根据NCBI数据库信息设计引物序列,采用PCR技术从抗虫棉基因组DNA中扩增出抗虫基因cry的保守序列,构建重组载体并转化到大肠杆菌Rosetta(DE3)菌株中,利用pGEX原核表达系统诱导蛋白表达。同时,分析了不同IPTG浓度及不同诱导时间对蛋白表达量的影响。[结果]扩增出抗虫基因cry的两段保守序列,长度分别为304和853 bp;SDS-PAGE检测结果显示,经IPTG诱导后重组质粒pGEX-4t-1-304和pGEX-4t-1-853成功地表达了大量GST融合蛋白,分子量分别为39和62.4 kDa,与预期结果一致;确定了IPTG最佳诱导浓度为0.15 mmol/L,最佳诱导时间为7h。[结论]为今后检测转Bt基因农作物奠定了基础。
罗翠平李金华谢烨明曾万勇
关键词:保守序列克隆
正交设计优化南苍术快速繁殖培养基的研究被引量:4
2012年
[目的]保护野生苍术资源,并为南苍术工厂化生产提供技术指导。[方法]以南苍术[Atractylodes lancea(Thunb.)DC]种子为材料得到无菌试管苗,应用正交试验法研究6-苄基腺嘌呤(6-BA)、萘乙酸(NAA)、激动素(KT)及其配比对南苍术无菌苗增殖的影响;同时研究了NAA浓度对南苍术苗生根的影响;以泥土、河沙、棉籽壳为材料,采用正交试验法设计9种轻型基质配方,研究其对南苍术生长的影响。[结果]适合南苍术试管苗增殖的培养基为MS+6-BA 1.0 mg/L+NAA 0.4 mg/L+KT 0.4 mg/L;适合南苍术生根的培养基为1/2MS+NAA 0.5 mg/L;最适基质配比为泥土∶河沙∶棉籽壳=3∶3∶2(体积比)。[结论]筛选出了南苍术增殖培养和生根的最佳配方,以及基质的最佳配比,为苍术资源的实验室保存和种苗工厂化生产提供了技术依据。
李文谢烨明罗翠平李金华曾万勇
关键词:正交设计激素基质
Cloning and Expression of Conserved Sequences of cry Gene in E.coli Strain Rosetta(DE3)被引量:1
2012年
[Objective] The aim was to clone the conserved sequences of cry gene and express them in Rosetta (DE3). [Method] Specific primers were designed according to NCBI database information and the conserved sequences of cry gene were amplified by PCR from Bt transgenic cotton. Then recombinant plasmids were constructed and expressed in E. coil strain Rosetta (DE3). Finally, the effects of different concentrations and inducing time of IPTG on the expression level of protein were investigated. [Result] Two conserved sequences (304 and 853 bp respectively) of cry gene were amplified. The result of SDS-PAGE confirmed that the recombinant plasmids pGEX-4t-I-304 and pGEX-4t-1-853 could express fusion proteins by IPTG induction and the molecular weight of protein products was 39 and 62.4 kDa respectively, which was in accordance with predicted result. The optimal protein ex- pression conditions were confirmed as induction with 0.15 mmol/L IPTG for 7 h. [Conclusion] This study prepared the ground for the further detection of Bt transgenic crops.
罗翠平李金华谢烨明曾万勇
关键词:CLONING
Study on Rapid Propagation Medium of Atractylodes Lances by Orthogonal Design被引量:1
2012年
[Objective] The aim was to protect wild resource of Atractylodes lances and to provide technical guidance for industrialization of Atractylodes lances. [Method] The test-tube seedlings were produced from seeds of Atractylodis Lances. At the same time, the effects of 6-BA, NAA and KT on the proliferation of aseptic seedling was studied through the orthogonal test. Effect of NAA concentration on the rooting of aseptic seedling was studied through nine types of light substrate groups, which consist of peat soil, fine sand and cotton seed hull, and their effects on Atractylodes lances growth were studied on with orthogonal design. [Result] The culture medium suitable for the proliferation of the Atractylodes lances tube seedling was MS+6-BA 1.0 mg/L+NAA 0.4 mg/L+KT 0,4 mg/L. And the medium suitable for rooting was 1/2MS+NAA 0.5 mg/L. The optimal ratio of soil, riversand and cotton seed hull was 3:3:2. [Conclusion] The research selected optimum culture medium, rooting medium and ratio of substrate, which provided technical guidance for laboratorial preservation of germplasm resource of Atractylodes lances and industrialization of its seedling production.
李文谢烨明罗翠平李金华曾万勇
关键词:HORMONESUBSTRATE
草胺膦浓度对黄芩植株生长和愈伤组织生长分化的影响被引量:1
2012年
通过研究除草剂草胺膦对黄芩(Scutellaria baicalensis Georgi)植株生长和愈伤组织生长分化的影响,发现草胺膦对黄芩植株生长和愈伤组织生长分化具有强烈的抑制作用。并在此基础上找到了合适的草胺膦筛选压,对于黄芩植株生长是30mg/L;对于黄芩愈伤组织生长是3.5mg/L;对于黄芩愈伤组织分化是3.0 mg/L。
罗翠平李金华杨玉稳曾万勇
关键词:愈伤组织除草剂
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